抗NPM1单克隆抗体的制备

目的 制备并鉴定NPM1单克隆抗体.方法 利用纯化的NPM1融合蛋白作为抗原,通过常规的细胞融合技术制备杂交瘤细胞,用间接ELISA法筛选能出产生抗NPM1单克隆抗体的阳性细胞株,对抗NPM1单克隆抗体的特性进行分析.结果 筛选出1株产生抗NPM1单克隆抗体的杂交瘤细胞株,产生的抗体为IgGl亚类,效价达1∶720000;纯化后经SDS-PAGE显示两条蛋白条带,纯度达95％以上,相对分子质量约为50000和25000,与单一抗体重链、轻链分子大小相符.免疫组化证实,纯化的2G7抗NPM1单克隆抗体可检测出NPM1在结直肠癌表达,阳性位点为细胞核.结论 制备的抗NPM1单克隆抗体可用于NPM1蛋白功能的研究与临床检测.

Preparation of the Monoclonal Antibody Against NPM1

Objective To prepare and identify the monoclonal antibody against NPM1 protein. Methods The purified NPM1 fusion protein was used as the antigen. The hybrid cells were prepared by the conventional cell fusion technology. The hybrid cell line produced monoelonal antibody against NPM1 was screened by indirect ELISA. The characterizations of this antibody were analyzed. Results A hybrid cell line that produces mono- clonal antibody against human NPM1 was screened. The subtype of antibody was IgG1 and the antibody titer reached 1 ： 720000 ; the purity of monoclonal antibody was determined by SDS- PAGE and showed the two bands correspond to single heave chain and light chain of an antibody. The IHC results also showed this could detect the NPM1 expression on colorectal cancer. Conclusion The monoclonal antibody against NPM1 prepared in this study could be used for NPM1 protein function research and clinical examination study.